CN112872002A - Application of selenium combined pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil - Google Patents
Application of selenium combined pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil Download PDFInfo
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Abstract
The invention discloses application of selenium combined pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil, belonging to the field of ecological environment protection. Lolium perenne is planted in cadmium-nonylphenol composite polluted soil, and a selenium source and Pseudomonas aeruginosa (Pseudomonas aeruginosa) are added to enhance the plant restoration effect. The selenium and the pseudomonas aeruginosa are combined with the ryegrass to treat the cadmium-nonylphenol composite contaminated soil, the selenium, the microorganisms and the plants have the effect of synergistically promoting pollution remediation, the ecological effect is good, and the method is suitable for remediation of the cadmium-nonylphenol composite contaminated soil. The method obviously improves the repairing efficiency of the ryegrass on the cadmium-nonylphenol composite polluted soil, and is simple to operate, low in cost and good in repairing effect.
Description
Technical Field
The invention relates to application of selenium combined pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil, belongs to the field of ecological environment protection, and particularly relates to the field of green remediation of heavy metal-organic matter composite contaminated soil.
Background
Nonylphenol (NP) is one of surfactant-degraded intermediate products widely used in industrial production, and is easily diffused into the soil environment along with surface runoff, so that soil nonylphenol pollution is caused. The soil nonylphenol pollution condition around the factory or by long-term wastewater irrigation is the most serious. Unreasonable human activities such as mining, industrial and agricultural production and the like cause the cadmium (Cd) pollution of soil to be increasingly serious, and the cadmium and nonyl phenol pollution in water, sludge and soil is serious in part of industrially developed areas. Environmental cadmium and nonyl phenol pollution cause quality and yield reduction of agricultural products, and can threaten human health through food chain or environmental exposure, which becomes an environmental problem to be solved urgently at present.
At present, methods for remedying the soil polluted by heavy metals and organic matters mainly comprise adsorbents, extractants, electrochemistry, bioremediation and the like, but most of the methods are difficult to popularize due to the problems of high cost, high technical requirements, large environmental disturbance and the like. The phytoremediation is a remediation method with low cost, environmental friendliness, no secondary pollution and strong sustainability. In single phytoremediation, due to the limitation of factors such as biomass, growth cycle and the like, cadmium and nonylphenol in soil are difficult to remove simultaneously and efficiently, so how to increase the removal rate of cadmium and nonylphenol in soil while increasing the biomass of plants becomes the key for phytoremediation of cadmium-nonylphenol composite contaminated soil.
The ryegrass has strong enrichment capacity on heavy metal elements such as Cd, Cu, Pb and the like, and the content of heavy metals in soil can be continuously reduced by cutting the ryegrass planted in the field; meanwhile, the ryegrass also has certain restoration capacity to organic pollution of soil Polycyclic Aromatic Hydrocarbons (PAHs) and the like, so the ryegrass is suitable for being used as a plant for restoring cadmium-nonylphenol composite polluted soil. The microorganism is a main participant of metabolic activity in a soil-plant system, can degrade organic matters through a specific enzyme way, can convert the occurrence form of heavy metal, or can repair heavy metal pollution through processes of extracellular complexation, intracellular accumulation and the like. In addition, part of the microorganisms have obvious effects on regulating soil nutrient circulation and promoting plant growth, namely, the absorption and accumulation amount of part of plants to pollutants can be improved. In addition to microorganisms, selenium is also commonly used for soil pollution remediation. Selenium is one of the beneficial elements of plants, can promote the growth of the plants, enhance the stress resistance of the plants, and even can react with heavy metals to generate a compound (such as a Cd-Se compound). In order to further improve the phytoremediation efficiency, microorganism reinforcement, trace element reinforcement and phytoremediation can be combined, the biomass of plants is improved, and the plants are promoted to absorb and accumulate pollutants and degrade organic pollutants.
Disclosure of Invention
Aiming at the practical problems and requirements in production practice, the invention aims to provide a method for strengthening plant repair of cadmium-nonylphenol composite contaminated soil by selenium combined with pseudomonas aeruginosa. Cadmium and nonyl phenol in the soil are removed by planting ryegrass, and green ecological restoration of the cadmium-nonyl phenol composite polluted soil is realized.
The invention discovers that the growth of ryegrass in cadmium-nonylphenol composite polluted soil can be promoted by singly inoculating Pseudomonas aeruginosa (Pseudomonas aeruginosa), singly applying selenium and jointly applying Pseudomonas aeruginosa and selenium, the absorption and accumulation of cadmium by ryegrass can be improved, cadmium and nonylphenol in the soil can be removed, and the treatment effect of the joint application of Pseudomonas aeruginosa and selenium is optimal. In addition, the invention also discovers that the catalase activity can be improved by adding selenium or simultaneously applying pseudomonas aeruginosa and selenium in the cadmium-nonylphenol composite contaminated soil. In conclusion, the selenium and pseudomonas aeruginosa are beneficial to strengthening the repairing effect of the plants on the cadmium-nonylphenol composite polluted soil.
Specifically, the invention provides the following technical scheme:
application of selenium combined with pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil.
The application of selenium combined pseudomonas aeruginosa in strengthening absorption of cadmium and nonylphenol in cadmium-nonylphenol composite polluted soil by plants.
The application of selenium combined with pseudomonas aeruginosa in improving the catalase activity of cadmium-nonylphenol composite contaminated soil.
A method for repairing cadmium-nonylphenol composite contaminated soil is to apply a selenium source and pseudomonas aeruginosa to the cadmium-nonylphenol composite contaminated soil and plant the selenium source and the pseudomonas aeruginosa.
A method for improving catalase activity of cadmium-nonylphenol composite contaminated soil is to apply a selenium source and pseudomonas aeruginosa in the soil.
Preferably, the selenium applying source and the pseudomonas aeruginosa are selenium applying solution and pseudomonas aeruginosa bacteria solution.
Preferably, the plant is ryegrass.
A composition for repairing cadmium-nonylphenol composite contaminated soil comprises a selenium source and pseudomonas aeruginosa.
A composition for removing cadmium and nonylphenol from cadmium-nonylphenol composite contaminated soil comprises a selenium source and pseudomonas aeruginosa.
A composition for promoting ryegrass to absorb cadmium and nonylphenol in cadmium-nonylphenol composite contaminated soil comprises a selenium source and pseudomonas aeruginosa.
A composition for improving catalase activity of cadmium-nonylphenol composite contaminated soil comprises a selenium source and pseudomonas aeruginosa.
The selenium source is preferably sodium selenate.
Compared with the prior art, the method for restoring the cadmium-nonylphenol composite polluted soil has the following advantages:
(1) the selenium and pseudomonas aeruginosa are combined to reinforce the restoration of the cadmium-nonylphenol composite contaminated soil by the plants, so that the cadmium and nonylphenol in the soil can be effectively removed, and the defects of the research on the green restoration technology of the heavy metal-organic matter composite contaminated (especially cadmium-nonylphenol composite contaminated) soil at home and abroad at the present stage are overcome;
(2) the selenium and pseudomonas aeruginosa are combined to strengthen the restoration of the cadmium-nonylphenol composite contaminated soil by the plants, so that the restoration efficiency of the cadmium-nonylphenol composite contaminated soil by the plants is improved, and the phenomenon of synergistically enhancing the restoration effect exists among microorganisms, trace elements and the plants;
(3) the application method of the selenium source and the bacterial liquid is simple, and the biomass of the ryegrass can be obviously increased by applying the selenium source and the bacterial liquid once, so that the activity of the soil enzyme is improved, the tolerance of the plant to the cadmium-nonylphenol composite pollution is improved, and the method is suitable for in-situ remediation of the cadmium-nonylphenol composite pollution soil.
Drawings
FIG. 1 is a graph of the results of cadmium content in the lower part of a ryegrass plant.
FIG. 2 is a graph of the results of the aerial cadmium content of Lolium perenne plants.
FIG. 3 is a result graph of cadmium content in cadmium-nonylphenol composite contaminated soil.
FIG. 4 is a graph showing the results of the content of nonylphenol in cadmium-nonylphenol complex contaminated soil.
FIG. 5 is a graph showing the results of catalase activity in cadmium-nonylphenol complex contaminated soil.
In the figure, different letters a, b, c, etc. indicate the significance of the difference between different treatments.
Detailed Description
The following examples are intended to further illustrate the invention but should not be construed as limiting it. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1: application of selenium-combined pseudomonas aeruginosa in strengthening plant remediation of cadmium-nonylphenol composite contaminated soil
The test site is a microelement central potted plant (30 degrees 28 '26' N and 114 degrees 02 '15' E) of Huazhong agriculture university, the soil to be tested is cadmium-polluted vegetable garden soil in Wuhan city, Hubei province, the basic physicochemical properties of the soil are that pH is 5.99 (soil-water ratio is 1:2.5), organic matter is 9.39g/kg, alkaline hydrolysis nitrogen is 41.07mg/kg, quick-acting phosphorus is 22.28mg/kg, quick-acting potassium is 160mg/kg, the total cadmium content is 1.48mg/kg, and nonyl phenol is not detected. And naturally drying the soil sample, grinding, sieving and storing for later use.
Cadmium-nonyl phenol composite contaminated soil: and the low-concentration nonyl phenol (5mg/kg soil) and the high-concentration nonyl phenol (50mg/kg soil) are respectively compounded with cadmium to pollute the soil, namely the high-and low-compound polluted soil. Respectively taking a small amount of cadmium-polluted soil, adding 5mg/mL of nonylphenol-ethanol stock solution into the cadmium-polluted soil, uniformly stirring, standing for 48 hours, mixing the mixture into other nonylphenol-free soil when ethanol is completely volatilized, wherein the total weight of the soil in each pot is 2.5kg, and uniformly stirring to ensure that the concentration of nonylphenol in the soil is respectively 5mg/kg and 50 mg/kg.
The ryegrass tested was a bindwood tetraploid.
The test strain is Pseudomonas aeruginosa (Pseudomonas aeruginosa) strain SH1 provided by Suzhou university of science and technology (Schwanguu et al, 2017, influence of cadmium on degradation of nonyl phenol in water by Pseudomonas aeruginosa).
Nutrient liquid culture medium: 5g of glucose, 2g of peptone and 1g of yeast powder are dissolved in 1000mL of deionized water, the pH value is adjusted to 7.2-7.5, and autoclaving is performed.
Solid medium: 5g of NaCl, 5g of beef extract, 10g of peptone and 20g of agar are dissolved in 1000mL of deionized water, the pH value is adjusted to 7.0-7.2, and the mixture is autoclaved.
Preparing pseudomonas aeruginosa bacterial liquid: selecting 1-ring pseudomonas aeruginosa from a solid culture medium, inoculating the pseudomonas aeruginosa into 100mL of nutrient liquid culture medium, and culturing at the temperature of 30 ℃ for 150 r.min-1Shaking and culturing for 24h at constant temperature. Inoculating the bacteria liquid into fresh nutrient culture solution according to the volume ratio of 1% by adopting an aseptic technique, and performing constant-temperature shake culture for 24 hours under the same condition. Suspending the bacteria at 6000r min-1Centrifuging for 10min, discarding supernatant to obtain thallus, washing with phosphate buffer, and adding sterile water to prepare bacteria solution.
The test was run with a total of eight treatments, four replicates per treatment, as shown in table 1. Exogenous selenium was applied as sodium selenate at a concentration of 1.0mg/kg soil.
TABLE 1 test treatment
The pot culture test was carried out using urea, sodium dihydrogen phosphate, and potassium chloride (N: 180 mg/kg; P)2O5：100mg/kg；K2O: 150mg/kg), etc., and applying 50mL of 0.12g/L Na according to the treatment2SeO4Solution, according to 108Inoculating pseudomonas aeruginosa bacterial liquid to the strain/g soil, and sowing the pseudomonas aeruginosa bacterial liquid after two days. The method comprises the steps of selecting full ryegrass seeds, drying the seeds in the sun for 1h, soaking the seeds in 10% NaClO for 30min, washing the seeds in tap water and deionized water for three times respectively, soaking the seeds in the deionized water for 12h, sowing 60 seeds in each pot, and covering a layer of thin soil on the surface of the seeds. Thinning is carried out within 7 days after germination until 30 ryegrass seedlings are planted in each pot, and ryegrass is cultured for 60 days, and is irrigated with deionized water. After 60 days of ryegrass growth, the ryegrass plant samples are harvested on the ground and underground. Storing part of fresh soil in a refrigerator at the temperature of-20 ℃ for subsequent soil enzyme activity determination, uniformly mixing other soil, taking part of the mixed soil by a quartering method, and air-drying and sieving the part of the mixed soil for determining the content of pollutants. Washing overground and underground parts of ryegrass with tap water and deionized water respectively, weighing, deactivating enzyme at 105 ℃ for 30 minutes, drying at 65 ℃ to constant weight, crushing, and sieving to be tested.
The effect of selenium in combination with pseudomonas aeruginosa on ryegrass biomass in cadmium-nonylphenol combined contaminated soil is shown in tables 2 and 3. As can be seen from the data in the table: the growth of ryegrass is promoted by singly inoculating pseudomonas aeruginosa in low-pollution soil or high-pollution soil, singly applying exogenous selenium and jointly applying bacteria and selenium, so that the biomass of ryegrass is improved, and the promotion effect of simultaneously applying bacteria and selenium is optimal.
TABLE 2 Lolium perenne biomass in soil compositely contaminated by low-concentration cadmium and nonyl phenol
TABLE 3 rye grass biomass of soil compositely contaminated by high-concentration cadmium and nonyl phenol
In the table, a, b and c represent the significance of the difference between the different treatments.
The cadmium content in the underground and overground parts of the ryegrass is measured by using a mixed acid (nitric acid: perchloric acid (v/v) ═ 9:1) digestion- (graphite furnace) atomic absorption spectrometer. The effect of selenium and pseudomonas aeruginosa on cadmium content in the underground and overground parts of ryegrass in the cadmium-nonylphenol composite polluted soil is shown in fig. 1 and fig. 2. The results show that the cadmium content of the underground part of the plant is improved by the three repairing measures, and the cadmium content of the overground part of the ryegrass can be obviously improved by singly applying exogenous selenium or simultaneously applying bacteria and selenium. Under the condition of low pollution, the strengthening effect of applying the bacteria and the selenium to the plant to repair the cadmium is the best. The combined application of selenium and bacteria increases the cadmium content of underground and overground parts of ryegrass by 4.1% and 49.1% respectively, compared with ryegrass grown in the soil compositely polluted without the application of bacteria or selenium. Under the high pollution condition, the cadmium content of the underground part of the plant is improved by 28.5 percent after the pseudomonas aeruginosa is inoculated independently compared with the cadmium content of the plant without the bacterium or selenium treatment; the selenium and the bacteria are applied together to promote the cadmium accumulation on the overground part of the ryegrass, and the cadmium content is 75.0 percent higher than that of the ryegrass without the bacteria or the selenium.
The effect of each repairing measure can be directly reflected by comparing the cadmium content in different soil samples. A soil sample is digested by a microwave digestion method, and then the total cadmium content of the soil is determined by a graphite furnace atomic absorption spectrometer, in particular to a graphite furnace atomic absorption spectrophotometry for determining lead and cadmium which are solid wastes of HJ 787-2016. The influence of exogenous selenium and pseudomonas aeruginosa on the cadmium content in the composite contaminated soil is shown in fig. 3. Under the conditions of high pollution and low pollution, the cadmium content of the soil after 60 days of culture is respectively 1.32mg/kg and 1.26mg/kg, after three repairing measures are applied, the cadmium content of the soil is obviously reduced, the effect of simultaneously applying bacteria and selenium is most obvious, under the conditions of high pollution and low pollution, the cadmium content of the soil is respectively 0.87mg/kg and 0.79mg/kg, and is respectively reduced by 34.1 percent and 36.9 percent compared with the soil without applying bacteria or selenium. The effect of the selenium and the bacteria combined application of the enhanced ryegrass for repairing cadmium in the cadmium-nonylphenol combined polluted soil is shown to be most obvious.
The soil nonylphenol content is one of the most important indexes for measuring the remediation effect, the soil nonylphenol content is measured by adopting a high performance liquid chromatography, a certain amount of anhydrous sodium sulfate is added into 2g of soil, the nonylphenol is extracted by using 5mL of an organic extracting agent (methanol: toluene (v/v) ═ 6:1), the ultrasonic treatment is carried out for 40min, and the extracting solution is collected into a nitrogen blow pipe. The above procedure was repeated four times in succession. Finally, the collected solvent was flushed to 1mL with nitrogen and analyzed by high performance liquid chromatography. High performance liquid chromatography was used, Fluorescence (FLR) detector, mobile phase methanol-water-acetonitrile (v/v/v ═ 70:20:10), flow rate 1mL/min, injection volume 20 μ L. The FLR excitation and emission wavelengths were 275nm and 308nm, respectively.
As shown in FIG. 4, the soil without applied bacteria or selenium still had a high nonylphenol content after 60 days of cultivation, and the nonylphenol contents of the high and low pollution level soils were 44.27mg/kg and 4.81mg/kg, respectively. The three restoration measures can reduce the concentration of the nonyl phenol in the soil, wherein the effect of singly inoculating the pseudomonas aeruginosa and the combined application of the selenium and the bacteria is obvious, the combined application treatment effect of the selenium and the bacteria is optimal, the nonyl phenol content in the soil with high and low pollution levels is respectively reduced to 15.71mg/kg and 1.02mg/kg, and the removal rate is as high as 78.8 percent and 62.8 percent. The pseudomonas aeruginosa is shown to be a main factor for removing the nonyl phenol in the soil, and when exogenous selenium and the bacteria are jointly applied, the selenium and the bacteria have a synergistic effect, so that the removal of the nonyl phenol can be further promoted.
The hydrogen peroxide is generated in the reaction processes of biological respiration process, biochemical oxidation and the like in the soil, the hydrogen peroxide has toxic action on plants and soil organisms, and the toxic action of heavy metals and organic pollutants aggravates the generation of the hydrogen peroxide and seriously threatens the environmental health of the soil and plants. Meanwhile, catalase exists in organisms and soil, so that the decomposition of hydrogen peroxide can be promoted, and the toxic action of the hydrogen peroxide is reduced. Therefore, the soil catalase activity can reflect the detoxification capability of the soil system. By adopting KMnO4The soil catalase activity was determined by titration, and 5g of the soil sample was weighed and placed in a 150mL triangular flask, into which 40mL of distilled water and 5mL of 0.3% hydrogen peroxide solution were introduced. Shaking on 120r/min shaking table for 30 min. Then 5mL of 1.5mol/L H was injected2SO4To terminate the reaction, filtration was carried out. Taking 25mL of filtrate, and adding 0.002mol/L of filtrateThe potassium manganate solution was titrated to reddish color while setting the control. Catalase activity was reflected in the volume of potassium permanganate consumed per unit soil weight.
The assay results (fig. 5) show that selenium in combination with bacteria significantly increased catalase activity under low-contamination conditions; under high pollution conditions, the catalase activity can be obviously improved by singly applying selenium and jointly applying selenium and bacteria. The combined application of selenium and bacteria has the most obvious effect on the activity of soil catalase, so that the catalase activity in the soil with high and low pollution levels is respectively improved by 35.8 percent and 38.1 percent. The selenium and the bacteria are applied together, so that the detoxification capability of cadmium-nonylphenol composite contaminated soil for ryegrass growth can be improved, the biological poisoning resistance of cadmium and nonylphenol for plants and soil is facilitated, and the phytoremediation effect is enhanced.
The foregoing embodiments are illustrative of the present invention, and various modifications can be made without departing from the spirit and scope of the invention. For example, the preparation method of the bacterial liquid, the concentration of cadmium and nonyl phenol, the culture time and the culture mode of ryegrass and the like are changed. And similar modifications are intended to be included within the scope of the present invention.
Claims (10)
1. The application of selenium combined with pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol combined contaminated soil is characterized in that: the plant is ryegrass.
2. The application of selenium combined pseudomonas aeruginosa in strengthening the absorption of cadmium and nonylphenol in cadmium-nonylphenol composite polluted soil by plants is characterized in that: the plant is ryegrass.
3. The application of selenium combined with pseudomonas aeruginosa in improving the catalase activity of cadmium-nonylphenol composite contaminated soil.
4. A method for repairing cadmium-nonylphenol composite contaminated soil is characterized by comprising the following steps: the method comprises the steps of applying a selenium source and pseudomonas aeruginosa in cadmium-nonylphenol composite polluted soil, and planting ryegrass.
5. A method for improving catalase activity of cadmium-nonylphenol composite contaminated soil is characterized by comprising the following steps: the method is characterized in that a selenium source and pseudomonas aeruginosa are applied in soil.
6. The composition for repairing cadmium-nonylphenol combined polluted soil is characterized by comprising the following components in parts by weight: comprises a selenium source and pseudomonas aeruginosa.
7. The composition for removing cadmium and nonyl phenol in cadmium-nonyl phenol combined polluted soil is characterized by comprising the following components in parts by weight: comprises a selenium source and pseudomonas aeruginosa.
8. The composition for promoting ryegrass to absorb cadmium and nonyl phenol in cadmium-nonyl phenol combined polluted soil is characterized by comprising the following components in parts by weight: comprises a selenium source and pseudomonas aeruginosa.
9. The composition for improving the catalase activity of the cadmium-nonylphenol composite contaminated soil is characterized by comprising the following components in parts by weight: comprises a selenium source and pseudomonas aeruginosa.
10. The method of claim 4 or 5 or the composition of any one of claims 6-9, wherein: the selenium source is sodium selenate.
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Application publication date: 20210601 |